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cDNA cloning and endogenous ligand identification of the novel ligand-gated ion-channel receptors

新型配体门控离子通道受体的 cDNA 克隆和内源配体鉴定

基本信息

批准号：
16500231
负责人：
HOUTANI Takeshi
金额：
$ 1.92万
依托单位：
Kansai Medical University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2004
资助国家：
日本
起止时间：
2004 至 2005
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16500231/
关键词：
serotonin ion channel human genome gene expression RT-PCR

项目摘要

5HT3C is the new member of serotonin-gated ionotropic receptor. From the sequence homology search for 5HT3C cDNA against human genomic database, three additional receptor genes, designated 5HT3D, 3E and 3L1, were identified on the same chromosome 3q27.1. These four predicted genes are composed of nine exons and share common features in exon-intron organization with previously known serotonin receptors 5HT3A and 3B. RT-PCR analysis in human tissues demonstrated that 5HT3C mRNA was highly expressed in placenta, lung and kidney, whereas 5HT3D mRNA was seen in lung and pancreas. The cDNA cloning revealed that alternative splicing form lacking exon 2,3,4 and 6 of 5HT3C and one lacking exon 3 of 5HT3D were expressed in kidney and pancreas, respectively. To compare the expression levels of these mRNA in each tissue, PCR-RFLP assay was performed by using common primers designed from the conserved sequences in exon 7 and exon 9. In brain, lung and liver, 5HT3C transcript was dominant, while 5HT3D message was mainly expressed in heart. Coexpression of 5HT3D and 5HT3L1 was demonstrated in skeletal muscle while 5HT3C, 5HT3D and 5HT3L1 were co-expressed in placenta, kidney and pancreas. The expression of 5HT3E transcript could not be detected, and hence, this gene was assumed as a pseudogene. Furthermore, other splicing variants at the junction of exon 7 and exon 8 of 5HT3D and 5HT3L1 were expressed in pancreas. Taken together, these results suggest that novel identified ion-channel receptors execute or modulate many physiological functions in diverse tissues through oligomerization of various splicing variants.

5 HT 3C是降钙素门控离子型受体的新成员。从针对人类基因组数据库的5 HT 3C cDNA的序列同源性搜索中，在同一染色体3q27.1上鉴定了三个另外的受体基因，命名为5 HT 3D、3E和3L 1。这四个预测的基因由九个外显子组成，并且与先前已知的5-羟色胺受体5 HT 3A和3B在外显子-内含子组织中具有共同特征。RT-PCR结果显示，5-HT-3C mRNA在胎盘、肺和肾组织中高表达，而5-HT-3D mRNA在肺和胰腺组织中高表达。cDNA克隆显示，缺失5 HT 3C第2、3、4、6外显子的选择性剪接体和缺失5 HT 3D第3外显子的选择性剪接体分别在肾脏和胰腺中表达。为了比较这些mRNA在每个组织中的表达水平，通过使用从外显子7和外显子9的保守序列设计的通用引物进行PCR-RFLP分析。在脑、肺和肝中，5 HT 3C转录占优势，而5 HT 3D信使主要在心脏中表达。在骨骼肌中发现5 HT 3D和5 HT 3L 1共表达，而在胎盘、肾脏和胰腺中发现5 HT 3C、5 HT 3D和5 HT 3L 1共表达。未检测到5 HT 3E转录本的表达，因此，该基因被认为是假基因。此外，5 HT 3D和5 HT 3L 1的外显子7和外显子8连接处的其他剪接变体在胰腺中表达。总之，这些结果表明，新的离子通道受体执行或调节多种生理功能，在不同的组织通过寡聚化的各种剪接变异体。

项目成果

期刊论文数量（20）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Enhanced hippocampal acetylcholine release in nociceptin-receptor knockout mice

DOI：
10.1016/j.brainres.2005.05.044
发表时间：
2005-07
期刊：
Brain Research
影响因子：
2.9
作者：
K. Uezu;A. Sano;H. Sei;K. Toida;T. Houtani;T. Sugimoto;Toshiko Suzuki-Yamamoto;H. Takeshima;K. Ishimura;Y. Morita
通讯作者：
K. Uezu;A. Sano;H. Sei;K. Toida;T. Houtani;T. Sugimoto;Toshiko Suzuki-Yamamoto;H. Takeshima;K. Ishimura;Y. Morita

Lack of nociceptin receptor alters body temperature during resting period in mice.

缺乏伤害感受肽受体会改变小鼠休息期间的体温。